An optimized fluorescent probe for visualizing glutamate neurotransmission

Jonathan S. Marvin, Bart G. Borghuis, Lin Tian, Joseph Cichon, Mark T. Harnett, Jasper Akerboom, Andrew Gordus, Sabine L. Renninger, Tsai Wen Chen, Cornelia I. Bargmann, Michael B. Orger, Eric R. Schreiter, Jonathan B. Demb, Wen Biao Gan, S. Andrew Hires, Loren L. Looger

Research output: Contribution to journalArticle

366 Scopus citations

Abstract

We describe an intensity-based glutamate-sensing fluorescent reporter (iGluSnFR) with signal-to-noise ratio and kinetics appropriate for in vivo imaging. We engineered iGluSnFR in vitro to maximize its fluorescence change, and we validated its utility for visualizing glutamate release by neurons and astrocytes in increasingly intact neurological systems. In hippocampal culture, iGluSnFR detected single field stimulus-evoked glutamate release events. In pyramidal neurons in acute brain slices, glutamate uncaging at single spines showed that iGluSnFR responds robustly and specifically to glutamate in situ, and responses correlate with voltage changes. In mouse retina, iGluSnFR-expressing neurons showed intact light-evoked excitatory currents, and the sensor revealed tonic glutamate signaling in response to light stimuli. In worms, glutamate signals preceded and predicted postsynaptic calcium transients. In zebrafish, iGluSnFR revealed spatial organization of direction-selective synaptic activity in the optic tectum. Finally, in mouse forelimb motor cortex, iGluSnFR expression in layer V pyramidal neurons revealed task-dependent single-spine activity during running.

Original languageEnglish (US)
Pages (from-to)162-170
Number of pages9
JournalNature Methods
Volume10
Issue number2
DOIs
StatePublished - Feb 2013

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Biochemistry
  • Cell Biology

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    Marvin, J. S., Borghuis, B. G., Tian, L., Cichon, J., Harnett, M. T., Akerboom, J., Gordus, A., Renninger, S. L., Chen, T. W., Bargmann, C. I., Orger, M. B., Schreiter, E. R., Demb, J. B., Gan, W. B., Hires, S. A., & Looger, L. L. (2013). An optimized fluorescent probe for visualizing glutamate neurotransmission. Nature Methods, 10(2), 162-170. https://doi.org/10.1038/nmeth.2333